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WO1997031622A1 - Stimulation de reponses immunitaires au moyen de faibles doses d'interleukine-2 - Google Patents

Stimulation de reponses immunitaires au moyen de faibles doses d'interleukine-2 Download PDF

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Publication number
WO1997031622A1
WO1997031622A1 PCT/US1997/003306 US9703306W WO9731622A1 WO 1997031622 A1 WO1997031622 A1 WO 1997031622A1 US 9703306 W US9703306 W US 9703306W WO 9731622 A1 WO9731622 A1 WO 9731622A1
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Prior art keywords
agent
composition
group
subject
cells
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PCT/US1997/003306
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English (en)
Inventor
Kendall A. Smith
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Cornell Research Foundation, Inc.
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Application filed by Cornell Research Foundation, Inc. filed Critical Cornell Research Foundation, Inc.
Priority to EP97916719A priority Critical patent/EP0893991A4/fr
Priority to AU25267/97A priority patent/AU2526797A/en
Publication of WO1997031622A1 publication Critical patent/WO1997031622A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/20—Interleukins [IL]
    • A61K38/2013—IL-2

Definitions

  • B cells In the immune system, there are three major types of lymphocytes: B cells, T cells, and natural killer (NK) cells.
  • B-cells are derived from bone marrow, and comprise about 10 % of the lymphocytes found circulating in blood. When stimulated by a specific antigen, each B-cell differentiates into a plasma cell that secretes antibodies of a single specificity. T-cells mature in the thymus and, make up about 80 % of circulating lymphocytes. Although not producing antibodies, T-cells bear on their surfaces specific antigen receptors resembling antibody molecules. T-cells react to antigen stimulation by secreting immunomediator molecules or cytokines (helper T- cells), and toxic molecules (cytotoxic T-cells).
  • Cytotoxic T-cells act directly on infected cells, and by secreting toxic molecules kill them and any foreign panicles, such as microorganisms, they may contain.
  • NK cells make up about 10% of the lymphocyte population, and are not antigen specific, but recognize and kill cells infected with microbes.
  • Monocytes and macrophages are large scavenger cells that ingest foreign particles and present antigens to the T- cells, which trigger specific immune responses. When an antigen is introduced, it is initially ingested by macrophages and other antigen presenting cells. After digestion, short segments thereof are presented on their cell surfaces. Only a few of all circulating T-cells have receptors that specifically bind to the antigen, and this binding stimulates the T-cells to secrete cytokines.
  • Cytokines are small proteins secreted primarily, but not exclusively, by cells of the immune system that promote the proliferation and/or differentiative functions of other cells.
  • cytokines include interleukins, interferons, hematopoietic colony stimulating factors (CSF), and proinfiammatory factors such as tumor necrosis factor (TNF).
  • Interleukin 2 IL-2
  • IL-2 acts on the three major types of lymphocytes, including T cells, B cells, and NK cells, stimulating them to proliferate and augmenting their differentiative functions.
  • IL-2 potentiates both innate or natural host defenses by stimulating NK cells, and antigen-specific acquired immune reactivity by stimulating T cells and B cells.
  • IL-2 mediates its effects by binding to specific receptors (IL-2R) expressed on the surfaces of IL-2-responsive target cells.
  • IL-2R specific receptors
  • IL-2R specific receptors
  • IL-2 binds to its receptors, it initiates a series of intraceliular events that result in the activation of a set of genes, which products determine the cellular responses observed, such as cellular proliferation.
  • IL-2 stimulation is an expansion in the number of T-cells, B-cells and NK cells.
  • IL-2 also stimulates the expression of genes encoding other cytokines, especially by T-cells and NK cells.
  • IL-2 stimulation of NK cells results in their production of secondary, pro-inflammatory cytokines, including TNFoc, interferon ⁇ (IFN), and GM-CSF.
  • cytokines including TNFoc, interferon ⁇ (IFN), and GM-CSF.
  • IFN interferon ⁇
  • GM-CSF GM-CSF
  • IL-2 has only been approved by the Food and Drug Administration for limited use in patients with renal cell carcinoma.
  • the high doses of IL-2 used only permit its administration for 1-2 weeks before severe toxicity develops. Additional rounds of treatment administered after discontinuance proved ineffective.
  • IL-2 units of biological activity have been converted to International Units (IU) to enable a comparison of the doses of IL-2 used in different studies.
  • An IU is defined as 15 x 10 6 units per mg I -2 protein.
  • some IL-2 doses are expressed as IU per m 2 of body surface area (BSA).
  • BSA body surface area
  • the BSA values are easily calculated from similar values based on a subject's weight and height using a standard conversion table of height and weight measurements. For example, a 70 kg. person of normal height has about 1.5 m 2 BSA. Mertlesman et al. administered daily subcutaneous doses of 1.5 to 3 x 10 5 IU impure, natural IL-2/m 2 to severely ill AIDS patients.
  • the administration was done by subcutaneous (s.q.) or intravenous (i.v.) injection for a short period of 3 weeks. Neither significant toxicities nor improvement in the measured immunologic parameters were reported.
  • rIL-2 recombinant IL-2
  • Rosenberg and co-workers were administered by Rosenberg and co-workers for the first time in 1985. Cancer patients were treated daily with high doses (up to 70 x 10 6 IU rIL-2/m 2 given in 3 divided i.v. bolus infusions 5 days/week for short periods of time (1-2 weeks). While this treatment was reported to yield anti-tumor responses in only about 20% of the patients, die dose and schedule of administration led to severe toxicities WHO Grades 3 and 4. (See, Table 3 below). In spite of the severe toxicity elicited, minor variations on these doses and schedules have also been used by others in the past 10 years for the treatment of many malignancies, as well as other indications because of its beneficial anti-tumor responses.
  • Caliguri and co-workers including the present inventor, first reported the administration of rIL-2 for a period longer man a few days or weeks to cancer patients.
  • IL-2 could be administered for as long as 3 months as a continuous i.v. infusion.
  • a daily dose of less than 0.45 x 10* IU rILr2/m 2 produced toxicities WHO grade 1
  • the highest dose tested, 0.6 x 10*IU rIL-2/m 2 /day produced toxicities WHO grade 2 and higher, requiring the lowering ofthe dose.
  • a dose of 0.45 x 10* IU rIL-2/m 2 /day produced a gradual increase in circulating NK cells over a 3-month interval.
  • concentration of circulating T-cells and B-cells failed to increase.
  • the same team (Soiffer et. al.) achieved similar results when treating hematological malignancies in bone marrow transplant patients by administration of a similar dose range and schedule.
  • Kaplan and co-workers including the present inventor, administered doses of rIL-2 in a range similar to that used by Caliguri et al. for only 8 days to patients with lepromatous leprosy. Although this treatment resulted in a 6-fold increase in circulating NK cells, it did not increase the concentration of circulating T cells and B cells. However, marked toxicity WHO Grade 1 was observed.
  • rIL-2 infusion via central or peripheral lines at a dose of 1-24 x 10* IU rIL-2/m 2 /day for 1 day to 2 weeks, preferably 5 days, with intervals between infusions of 4 weeks to 6 months.
  • these doses of rIL-2 are reported to cause severe toxic side effects, including capillary leak with hypotension, high fever (> 39°C), and extreme flu-like symptoms, necessitating hospitalization.
  • Example 8 is a paper example which forecasts the oral administration of ddl in combination with an all encompassing dose of IL-2 (25,000-1,000,000 U IL-2/day) by continuous infusion or other systemic administration for a period of 3 months. It is not indicated that these are IU. Although the example indicates that "beneficial results are seen" but, because it is entirely hypothetical, it provides no real information on IL-2 toxicity since no enabling work was provided.
  • This invention relates to a unit dosage composition, comprising in a sterile container, about 15,000 to 1,000,000 IU of an agent having IL-2 activity, including natural, recombinant and mutated IL-2, analogs and derivatives of IL-2, and to a kit provided with multiple unit dosages of the composition, instructions, and one or more devices for its administration, such as needles and syringes, inhalators, and the like.
  • composition may be in various forms, such as creams, ointments, sprays, solutions, suppositories, powders, suspensions, patches, emulsions, and encapsulated particles, among others, and may contain other therapeutic agents, such as additional lymphokines or cytokines, analogues, fragments, fusion proteins, and derivatives thereof, other biologically active agents and formulation additives.
  • Topical and controlled release formulations, implants, inhalators, and transdermal, transbuccal, and transpulmonary delivery devices are also included.
  • the agent may be self-administered by any topical or systemic route, as long as its peptide bonds are protected from degradation.
  • the present agent is suitable for the chronic stimulation and/or maintenance of immune response in a subject when administered at a dose effective to activate high affinity IL-2 receptors without eliciting substantial toxicity, i. e., WHO Group 1 or higher.
  • the amount of the agent administered may be determined as described herein, and is preferably effective to produce about 10 to about 90 % saturation of the high affinity IL-2 receptors.
  • a daily dose generally falls in the range of about 15,000 to 1,000,000 IU/m 2 .
  • the present immunotherapy is useful for stimulating the immune system in the prevention and treatment of malignancies, of mild and severe infections afflicting normal individuals, of opportunistic infections generally arising in immunocompromised individuals, such as viral, fungal, parasitic, and bacterial infections, including patients infected with the human immunodeficiency virus (HIV), and infections frequently encountered after operations, such as bone marrow transplants (BMT).
  • the present immunotherapy is also generally suitable for the stimulation of the immune system in normal subjects, and for use in animals in general.
  • FIGURE Figure 1 shows the IL-2 plasma concentration (Mean Standard Error) of samples from 6 subjects who received a daily dose of 125,000 IU recombinant IL-2/m 2 (8.3 ⁇ g/n )(•), and 8 subjects who received 250,000 IU recombinant IL-2/m 2 (16.7 ⁇ g/m 2 )( ), assayed at various times over a 24 hour period.
  • IL-2 plasma concentration Mean Standard Error
  • the present invention arose from a desire by the inventor to improve on prior art technology.
  • short term EL-2 therapy had been attempted with HIV+ patients, with patients suffering from some forms of cancer, and even with patients that had been subjected to bone marrow transplant (BMT).
  • BMT bone marrow transplant
  • high doses of IL-2 were shown to bring about extremely severe side effects, such as those classified by the World Health Organization (WHO) as Grades 1 to Grade 4 toxicities in Table 2 below.
  • WHO World Health Organization
  • IL-2 activity such as natural and recombinant IL-2, active analogues and derivatives of IL-2, or mixtures thereof, may be continousiy administered to patients for prolonged periods of time to activate and/or stimulate their immune system in the substantial absence of toxicity.
  • agents having IL-2 activity such as natural and recombinant IL-2, active analogues and derivatives of IL-2, or mixtures thereof.
  • the narrative will generically refer to IL-2, but the present teachings, unless otherwise indicated, extend to the remainder of the agents encompassed herein.
  • the effects of the present agents at the IL-2 receptors have been found to be dose-dependent, and to correlate to their binding affinity to the three different receptor classes expressed by different types of cells.
  • the high affinity class comprised of three peptide chains ( ⁇ , ⁇ , ⁇ ), is expressed by antigen-activated T-cells and B-cells (about 10 7 cells in d e circulation), and by about 10% of NK cells (total circulating about 8 10 cells).
  • This receptor class is characterized by a very high affinity for IL-2, as expressed by an equilibrium dissociation constant (Kd) of about 10 "u M.
  • IL-2 concentrations of about 10 "12 M to 10 10 M will bind to this class of IL-2R to different degrees.
  • a second class of functional IL-2Rs is comprised of only ⁇ and ⁇ receptor chains, and has a 100-fold lower affinity for IL- 2 than the high affinity receptor.
  • This intermediate affinity receptor has a Kd of about 10 "9 M, and is expressed by the majority (>90%) of NK cells (about 10 9 cells).
  • IL-2 concentrations which are expected to bind this class of receptors range from about 10 "10 to 10 "8 M.
  • the third class, a low affinity receptor comprises only ⁇ peptide chains, and has the lowest af inity for IL-2, with a Kd about 10 "8 M.
  • die EL-2 concentrations which are expected to bind to this class of receptor range from about 10* M to 10 "7 M.
  • target cells that express only ⁇ chains have not been identified, ⁇ chains have been detected in the serum as soluble IL-2Rs.
  • the soluble low affinity IL-2Rs cannot compete effectively with the cellular high affinity IL-2Rs.
  • the concentration of IL-2, the density of IL-2 receptors on the cell surface, and die duration of the IL-2-receptor interaction are three important parameters in the IL-2 regulation of lymphocyte proliferation and differentiation.
  • a finite number of IL-2-receptor interactions must occur before a cell irrevocably conunits to responding by proliferating. In normal individuals the vast majority of circulating T and B lymphocytes, i.e., greater than 99.9%, do not express IL-2Rs and are, thus, IL-2 unresponsive.
  • NK cells express IL-2Rs consitutively, with less than about 10% of NK cells (about 10* cells) expressing high affinity IL-2RS and > 90% of circulating NK cells (about 10 9 cells) expressing intermediate affinity IL-2 Rs.
  • the present inventor has shown that over a prolonged period of time, blood concentrations of about 10 "12 to 10 10 M IL-2 binding to high affinity IL-2Rs are therapeutic, whereas blood IL-2 concentration in excess of about 10 ⁇ 10 M, binding to and activating intermediate affinity IL-2Rs expressed by as many as about 10 9 NK cells, are toxic.
  • suitable doses for all agents in accordance with this invention may be calculated as follows.
  • the WHO grades for the toxicities produced by IL-2 were estimated from the above calculations, in accordance with this invention, and are shown in Table 3 below. They are based upon the World Health Organization (WHO) toxicity grading scale, shown in Table 2 above. Similar estimates may be done as taught herein for all the other agents of this invention.
  • XabkJ Prediction of IL-2 Toxicities
  • the severity of the toxicity is directly related to the blood concentration of IL-2.
  • the latter thus, together with the Kd value for the ligand-receptor interaction, may be used to calculate the degree of receptor occupancy as described above. If the Kd for the specific ligand-receptor is not known, it may be determined as described above or by other methods known in the art. Thereafter, using the above table, and having measured the peak plasma IL-2 concentration occurring, for example, about 2 hours after a subcutaneous injection, the practitioner may adjust, for example, the dosage of IL-2 for each individual subject.
  • a given dose yields a peak plasma concentration of 2xl0 '10 M
  • this concentration will lead to about 95% occupancy of high affinity IL-2 receptors, and more ti an about 16% intermediate affinity receptors.
  • the dose, dierefore, should be reduced gradually to that expected to saturate less than about 90% high affinity IL-2 receptors and less than about 10% intermediate affinity IL-2 receptors.
  • the ligand-receptor binding assay may be conducted, for the different agents in accordance with this invention, as described by Robb et al. (Robb, R., Munck, A., and Smith, K.A., J. Exp. Med. 154: 1455-1474 (1981)).
  • IL-2 or other ligands must be in their native configuration, and substantially free of contamination by other molecules.
  • the ligand may be labeled, for example, using radioisotopes, enzymes, and other markers.
  • Target cells isolated membranes, cytoplasm, or nuclei
  • Target cells may then be mixed together with the labeled ligand, and the ligand and receptor allowed to reach a steady state, where the rate of association of the ligand with the receptor about equals the rate of dissociation of the ligand from the receptor.
  • any unbound ligand may be separated from the receptor-bound ligand, usually by centrifuging the bound ligand, and the amounts of the bound and unbound ligand measured, e.g., as described by Robb et al., J. Exp. Med. 154: 1455-1474 (1981).
  • the affinity of the ligand- receptor interaction may be calculated, as well as the number of receptors per cell, or per weight of cytoplasm or nuclei, from these two experimentally determined values, knowing the number of cells, or amount of membranes, cytoplasm, or nuclei, used.
  • the data may be plotted by the method described by Scatchard (Ann. NY Acad. Sci. 51: 660-672 (1949).
  • the ratio of bound vs. free ligand may be plotted on the y-axis and the amount of bound ligand on the x axis.
  • the slope of the data points yields the Kd (x/y), whereas the x-axis intercepts yields the number of receptors. Knowing the equilibrium dissociation constant (Kd), the % receptors that will be occupied at each ligand concentration may be calculated as described above.
  • agent prescribed herein activates the immune system in a patient, but is non-toxic, so mat the subject may carry out normal daily activities.
  • asymptomatic HIV individuals have been shown by the inventor to be able to self-administer daily doses of agent in accordance with this invention safely and without toxicity for prolonged periods of time.
  • the present immunotherapy has been shown effective and free of toxicity WHO grade 1 or greater for a period greater than six months without the occurrence of systemic side effects or significant laboratory abnormalities.
  • an optimal dose range of a preparation of a specific recombinant IL-2 given to HTV+ individuals was determined to be below 500,000 IU/m 2 /day.
  • the present immunotherapy may be extended for one or two years, or longer periods, or be administered continuously as a maintenance therapy.
  • Some of the toxicities monitored to determine a suitable dosage for the recombinant IL-2 utilized in the reported study include weakness, fatigue, lethargy, myalgia, and low grade fever. These symptoms usually occurred within a few days after beginning the recombinant IL-2 injections.
  • the problem of toxicity observed at high IL-2 doses was circumvented by stopping therapy. The patients were then placed on a lower daily IL-2 dose, which proved safe. No toxicities to major organ systems were detected by laboratory assays throughout the full period of the test reported in this patent. All patients were, therefore, able to self-administer IL-2 daily, and perform normal activities without any hindrance associated with IL-2 side effects.
  • the low dose IL-2 administered in accordance with this invention does not trigger a burst in viral production of HIV reported previously upon the i.v. administration of high dose of IL-2.
  • increases as high as 50-fold persisting in plasma HIV levels were reported after a daily administration of 4 x 10* to 12 x 10* IU/m 2 IL-2 for 5 days.
  • an increase in viral production of that magnitude is due to the presence of pro-inflammatory cytokines, which are also responsible for the toxic symptoms observed with high dose IL-2 administration.
  • the present immunotherapy utilizes approximately 50-fold lower daily doses of IL-2, which do not result in a significant increase in plasma viral levels when compared with pre-treatment levels. Further yet, no opportunistic infections or malignancies are observed in any of the individuals receiving low doses of IL-2.
  • IL-2 utilized herein, when administered on a daily basis, stimulate the immune system, and significantly increase the number of circulating NK cells, eosinophils, monocytes, and CD4+ T cells.
  • high doses of IL-2 were administered by others as described above, i.v. intermittently every 2 months, it produced only transient increases in CD4+ T cells after each injection and the number of circulating NK cells, eosinophils and monocytes remained unchanged.
  • NK cells One group of cells believed to be critical to the overall immune response, the NK cells, were stimulated by the present method.
  • the number of NK cells increased, on the average, about 7-fold, from 67 ⁇ 12 cells/mm 3 prior to therapy to 464 ⁇ 187 cells/mm after 6 months of IL-2 therapy in accordance with the invention.
  • the total number of NK cells circulating in 5 1 of blood increased from 3.4 x If to 2.3 x 10 9 cells.
  • NK cells When NK cells are stimulated in vitro with IL-2, they become increasingly cytotoxic, and secrete cytokines that are potent stimuli for monocytes and macrophages.
  • the IL- 2-promoted accumulation of circulating NK cells increases their cytolytic capacity and the release of secondary cytokines important in the recruiting and activation of monocytes and macrophages.
  • the latter cells play a prominent role in an individual's natural (innate) immune responses.
  • the augmentation of antigen processing and presentation by monocytes and macrophages, promoted by NK cell-derived IFN- ⁇ , can augment antigen-specific, acquired immunoreactivity. Therefore, the increased number of circulating NK cells and monocytes elicited by die present immunotherapy contribute to improve both innate and acquired host defenses.
  • the daily administration of IL-2 in accordance with the invention also elicited a progressive accumulation of circulating CD4+ T-cells in all individuals, with a mean monthly gain of about 26 cells/mm 3 blood.
  • the intermittent administration of high doses of IL-2 was reported in the literature to lead to only a transient increase in CD4+ T-cell counts in 6 out of 10 subjects.
  • an increase of greater than about 300 cells is likely in the CD4+ T-cell count throughout a whole year of therapy.
  • the present invention provides a novel and effective method of stimulating the immune system of a subject by adntir ⁇ stering to the patient a daily dose of an agent having IL-2 activity, such as natural IL-2, an agent having active analogues, and derivatives thereof, which is effective to activate high affinity IL-2 receptors without eliciting substantial toxicity WHO grade 1 or higher.
  • an agent having IL-2 activity such as natural IL-2, an agent having active analogues, and derivatives thereof, which is effective to activate high affinity IL-2 receptors without eliciting substantial toxicity WHO grade 1 or higher.
  • compositions are provided herein for the administration of the agent of the invention under the conditions prescribed herein.
  • One of them is a topical composition, which in addition to the agent comprises a carrier or diluent for the agent, which is suitable for its transdermal delivery and optionally one or more of a variety of agents suitable for the preparation of different formulations, which will be selected in accordance with the type of formulation and route of administration desired.
  • ingredients are buffers, salt forming acids and bases, perfumes, colorants, emollients, adjuvants, single or multiple enteric coatings, copolymers, microporous or semi-permeable membranes, enzyme inhibitors, mucoadhesives, chelating agents, particulate systems, viral envelope proteins, liposomes and other micelles, emulsifiers, lipoproteins and other fatty acid derivatives, surfactants, bile salts, hydrophilic, neutral, and hydrophobic polymers and co-polymers, hydrogels, biodegradable polymers and co-polymers.
  • composition may also contain additional bioactive agents such as additional lymphokines or cytokines, and active analogues, fragments, fusion proteins, and pharmaceutically acceptable derivatives thereof, anti-bacterial, anti-viral, anti-fungal, anti-parasitic, anti-metabolic, anti- inflamrnatory, vasoactive, anti-neoplastic, bronchodilating, local anesthetic, immunomodulating, growth promoting and regenerating agents, enzymatic, hormonal agents, neurotransmitters, and cell receptor proteins and ligands.
  • additional lymphokines or cytokines and active analogues, fragments, fusion proteins, and pharmaceutically acceptable derivatives thereof, anti-bacterial, anti-viral, anti-fungal, anti-parasitic, anti-metabolic, anti- inflamrnatory, vasoactive, anti-neoplastic, bronchodilating, local anesthetic, immunomodulating, growth promoting and regenerating agents, enzymatic, hormonal agents, neurotransmitters
  • This composition may be in the form of a cream, an ointment, a solution, a gel, a powder, a suspension, an emulsion, encapsulated particles, or mixtures or combinations of these forms.
  • the agent may be present in different amounts, typically the dermal composition has about 0.001 to 50 wt% or more, and preferably about 0.1 to 30 wt% . However, other amounts larger and smaller are also suitable.
  • Another preferred form of die composition is in the form of a controlled release composition wherein the formulation ingredients added control the rate of release of die agent. These may be degradable polymers and copolymers, matrixes which "leach out" die agent, and the like, as is known in the art.
  • the composition may also be produced in die form of an implant for releasing a desired amount of the agent over a pre-determined period of time.
  • the composition of the invention may be provided as a kit with instructions for its use, particularly in terms of the any necessary manipulations, the number and timing ofthe applications, and the area to be applied to, as well as the frequency taking into consideration different body surface area and weight of the subject. This is particularly important when it is applied or administered to children, and more importantly to infants and newborn babies, as well as to small animals. Smaller doses may be required in these cases.
  • the composition may be delivered from a passive transdermal delivery device formed from a solid support with a compartment containing a solution or suspension comprising die composition.
  • the compartment has a permeable side which is applied to an area of a subject's skin or dermis and the agent is allowed to pass from the device onto and through the skin, mucosal, or buccal surfaces of d e subject.
  • the device is preferably placed in a sealed sterile container immediately after manufacture by methods known in the art.
  • a removable cover may be placed on the permeable side of the container prior to sealing and/or packaging to retain the solution or suspension of d e agent during storage and prior to use.
  • This device may be in the form of an electric transport device which also contains donor and counter electrodes, an external power source, and control circuitry.
  • This device may also be in the form of an ultrasound device which also contains an ultrasound generator (transducer), an external power source, and control circuitry.
  • ultrasound device which also contains an ultrasound generator (transducer), an external power source, and control circuitry.
  • Such devices are known in the art and need not be further described herein.
  • the solution or suspension containing the agent requires die presence of electric conducting agents which will aid in the transport of d e electrical current which facilitates the passage of the agent from the solution onto the skin or dermis and through it.
  • protein such as the present agent, it becomes important to provide means of administration that will not degrade or hydrolyze die peptic bonds, in order to preserve the activity of the agent.
  • die topical application of the agent in the form of a multiplicity of formulations is ideally suited to avoid the hydrolyzing conditions of die gastrointestinal tract while providing a direct passage, after the agent is absorbed through the skin or mucous tissue, directly into he blood stream.
  • Anodier particularly useful form of die invention is a unit dosage composition, which may be packaged in a sterile container, having about 15,000 to 1 ,500,000 IU, preferably up to about 1 ,000,000 IU, more preferably up to about 750,000 IU, and still more preferably up to about 500,000 IU, and as low as about 20,000 IU, preferably as low as about 50,000 IU, and still more preferably as low as about 60,000 IU of die agent of die invention, such as IL-2, pharmaceutically acceptable analogues and derivatives tiiereof, or mixtures tiiereof.
  • die agent of die invention such as IL-2, pharmaceutically acceptable analogues and derivatives tiiereof, or mixtures tiiereof.
  • the unit dosage composition may be prepared in any form to deliver die agent. Examples are powder, tablet, capsule, dragee, cream, solution, suspension, emulsion, gel, spray, or liposomal or other mi cellar forms. Preferred are solid, particularly freeze-dried, and liquid forms. Also preferred are other forms such as those suitable for injection, topical application, controlled release products, inhalation, and others.
  • Controlled release products are transdermal and intradermal devices, slow release oral formulations, patches, skin, mucosal and transbuccal implants, suppositories, and the like. Implants are preferred for long term delivery of the agent.
  • Controlled release products may be prepared as is known in die art and designed for releasing desired amounts of d e agent over a pre-determined period of time. In most instances, the amount ofthe agent contained in the control release product is substantially higher than die desired daily dose. In some cases, the product may contain sufficient amounts of agent for releasing a daily dose over a period of days, weeks, months, and even years. The product may be tested, and the amount of agent to be released adjusted in accordance with die observed absorbed dose.
  • the daily dose administered is preferably about 15,000 to up to about 1,000,000 IU/m 2 body surface.
  • the upper daily dose limit is preferably not higher than about 900,000 IU/m 2 , still more preferably not higher than about 800,000 IU/m 2 , and even more preferably not higher than 450,000 IU/m 2 .
  • the lower limit for the daily dose is preferably no lower tiian about 30,000 IU/m 2 , more preferably no lower than about 45,000 IU/m 2 , and even more preferably not lower than about 100,000 IU/m 2 .
  • the slow release of higher or lower amounts of the agent are also contemplated herein.
  • Any of die forms for administration of die agent may also contain other formulation components and additional bioactive agents as described herein.
  • the agent of this invention may also be administered from an inhalant device, including those utilized to introduce tiierapeutic or preventative agents into die lungs, as is the case of transpulmonary ad ⁇ iinistration to asthmatic patients.
  • an inhalant device including those utilized to introduce tiierapeutic or preventative agents into die lungs, as is the case of transpulmonary ad ⁇ iinistration to asthmatic patients.
  • Such devices are known in die art and available commercially. All types of products and compositions which are part of this invention may be provided as a kit, and preferably as a self-administration kit, primarily comprising, in separate sterile containers, a number of unit dosage compositions, and detailed instructions for use of the kit, and optionally, one or more devices including syringes and needles, transdermal, transbuccal, or intradermal patches or implants, or inhalators, for delivering the agent, and a carrier diluent. These devices may be placed in sterile containers, and die entire kit may
  • compositions and products provided herein may all be utilized for stimulating and thereafter rnamtaining the stimulation of the immune system of a mammalian animal by chronically administering the agent of die invention in die form of a composition over a prolonged period of time in an amount effective to activate the high affinity IL-2 receptors witiiout eliciting any substantial toxicity WHO Grade 1 or higher.
  • the present inventor found a manner to stimulate and maintain the stimulation of the immune system of a mammalian animal or a human without interfering with his/her normal activities or producing undesirable side effects which lower die quality of life of the individual.
  • the present immunotherapy may be administered for six months, one year, and for periods in excess of these times as long as parameters, including systemic symptoms of fatigue, malaise, myalgia, fever, and other symptoms typical of overstimulation of intermediate and low affinity IL-2 receptors shown in Table 1 above, are monitored.
  • die amount of agent administered should be effective to produce about 10 to 90 % saturation, preferably about 20 to 85% , more perfectly about 30 to 80% , and still more preferably about 50 to 70%, and in some cases below about 60% of the high affinity IL-2 receptors.
  • no toxicities are elicited even at agent occupancy greater than about 93 % , and even 95 %.
  • compositions may be administered by topical and systemic routes, including subcutaneous, intramuscular, intradermal, intralymphatic, mtratumor, transdermal, intradermal, intracavitary, oral, intranasal, intravaginal, intraa al, intrabuccal, transbuccal, transpulmonary, or subiingual routes, or by inhalation or implant.
  • die implants may be placed in die following areas: the limbs, thorax, abdomen and back, and die topical application, particularly when administered by means of a device, may be typically applied to the same areas.
  • the present immunotherapy may be applied as a preventative or therapeutic treatment to a variety of conditions.
  • IL-2 or any of die otiier agents disclosed herein having IL-2 activity been utilized to boost the immune system of normal animals or humans, including children.
  • the present technology may be utilized in humans as well as all types of mammalian animals, including equines, bovines, ovines, large and small animals in die care of a veterinarian, and animals found in die wild, among others.
  • die present invention extends to die preventative administration of die agent to normal subjects, for example in cases of travel to areas of the country or the world affording die possibility of a contagious infection, and even in die case of the common cold or flu that afflicts many people during die winter months.
  • the present immunotiierapy is also applicable to the prevention and treatment of viral, bacterial, fungal and parasitic infections in general, congenital or acquired immunodeficiencies and cancers or other malignancies such as carcinomas, melanomas, sarcomas, leukemias, ly phomas, and myelomas, among otiiers.
  • immunotiierapy witii the agent of the invention prevents increases in malignant cell mass or count, and in some instances, it even produces a reduction in tumor mass.
  • Other parameters that may be monitored in order to adjust the dose of the agent administered daily are the blood cell count of circulating lymphocytes, monocytes and poiymorphonuclear leukocytes, such as T-cells, B-cells, NK cells, monocytes, eosinophils, neutrophils, basophils, antigen-presenting cells, among others.
  • T-cells circulating lymphocytes
  • B-cells a cell that may be monitored in order to adjust the dose of the agent administered daily
  • NK cells such as T-cells, B-cells, NK cells, monocytes, eosinophils, neutrophils, basophils, antigen-presenting cells, among others.
  • T-cells T-cells
  • B-cells NK cells
  • monocytes eosinophils
  • neutrophils neutrophils
  • basophils basophils
  • antigen-presenting cells antigen-presenting cells
  • the present immunotiierapy does not increase die count of circulating virus or other opportunistic microorganisms when administered over a prolonged period of time, and in some cases it even decreases their count.
  • the immunotherapy is applied to subjects afflicted witii cancer, it is useful to monitor the size of the malignant cell mass or count.
  • This therapy may be administered as the main cancer therapy in the indicated types of malignancies or after surgery, chemotherapy or radiotherapy.
  • the present immunotherapy may encompass, in addition to the agent, nucleic acid analogues, other anti ⁇ viral drugs, and enzyme inhibitors, in amounts known in die art.
  • formulations containing one or more anti-viral agents of the nucleic acid analogue class and one or more protease inhibitors are particularly preferred.
  • the agent may be administered in die same formulation with these agents or by a different route. Different drugs may be alternated while die agent of the invention is administered over a prolonged period of time.
  • die combined immunotherapy in accordance witii tiiis invention should be started by lowering die dose of each individual drug and then progressively increasing their doses one at a time while monitoring the effect produced on one of more of the parameters followed, as described above.
  • An artisan with average skill in the art of medicine or veterinary science would know how to proceed to attain a desirable combination dose.
  • die agent may be self-administered by die human subject, preferably subcutaneously, transdermally, intrapulmonarily, transbuccally, or by implant.
  • die human subject preferably subcutaneously, transdermally, intrapulmonarily, transbuccally, or by implant.
  • present treatment is suitable for application to a variety of conditions, which benefit from the stimulation of the immune system.
  • the administration of low doses of die agent of this invention produces an increase in one or more of a group of circulating immunity-building cells, such as lymphocytes, monocytes and polymorphonuclear lymphocytes, including T-cells, particularly CD4+ T-cells, B-cells, natural killer (NK), eosinophils, monocytes, basophils, and antigen-presenting cells.
  • T-cells particularly CD4+ T-cells
  • B-cells B-cells
  • natural killer (NK) natural killer
  • eosinophils monocytes
  • basophils basophils
  • antigen-presenting cells antigen-presenting cells.
  • IL-2 One specific type of recombinant IL-2 has now been shown, in a clinical trial described below, to enhance the concentration of, for example, circulating NK cells, monocytes, eosinophils and CD4+ T-cells.
  • Suitable anti-viral agents are any of the agents utilized in the treatment of viral infections. Examples are zidovudine (AZT), 2',3'-dideoxyinosine (ddl), 3'-azido- 2', 3'-dideoxyd ⁇ ymidine, acyclovir, 1, 3- dihydro- 2 -propoxy-metiiyquanine (gancyclovir), ribavirin, dideoxycytidine (ddC), lamivudine (3TC), enzyme inhibitors, such as protease inhibitors, e. g., saquinovir, and invarase, among others, and combinations thereof.
  • ZTT zidovudine
  • ddl 2',3'-dideoxyinosine
  • ddC dideoxycytidine
  • ddC dideoxycytidine
  • lamivudine 3TC
  • enzyme inhibitors such as protease inhibitors, e. g.,
  • the anti-bacterial agent may be any one agent known to be effective against the agent associated with the infection. Examples are pentamidines, trimedioprim-sulfamedioxazole, sulfonamides, penicillins, cephalosporins, aminoglycosides, tetracyclines, chloramphenicols, and combinations thereof. Any specifically targeted antibodies and their fragments are suitable for use herein to deliver molecules attached to diem to specific sites. Examples are radioisotope, enzyme, toxin, and odier tiierapeutic agent-carrying monoclonal antibodies, and combinations thereof.
  • Any anti-fungal agent is suitable for use herein.
  • flucytosine amphotericin B, fluconazole, griseofulvine, and combinations thereof.
  • Any anti-parasitic agent is suitable for use herein.
  • examples are pyrimethamine, quinacrine, thiabendazole, levamisol, and combinations thereof.
  • Any anti-metabolic agent may be utilized herein.
  • Examples are purine analogues, folic acid analogues, pyrimidine analogues, and combinations thereof.
  • Any anu-irifkrrimatory agent is suitable for this invention.
  • examples are steroidal anti-inflammatory agents, non-steroidal anti-inflammatory agents such as acetaminophen, aspirin, and combinations thereof.
  • vasoactive agents such as epinephrine, norepinephrine, dopamine and combinations thereof, bronchodilating agents such as ⁇ 2 receptor agonists, and combinations thereof, local anesthetic agents such as procaine, cocaine, and combinations tiiereof, growth promoting and regenerating agents such as epidermal growth factor, fibroblast growth factor, and combinations thereof, additional lymphokines or cytokines such as interleukins other than IL-2, interferons, and the like, hematopoietins, growth factors, hormones, chemokines, active analogues, fragments, fusion proteins and pharmaceutically-acceptable derivatives thereof, and combinations thereof, agents such as soluble CD4 and analogues thereof, anionic polysaccharides, and anti-neoplastic agents such as alkylating agents, anti-metabolites, hormones, vinca, alkaloids, anti-proliferative agents, and combinations thereof.
  • Recombinant human IL-2 (R-Met-HU IL-2 (Ala 125 or rIL-2) was obtained from Amgen (Thousand Oaks, CA). Each 2 ml vial contained approximately 0.4 mg/ml of recombinant IL-2 (rIL-2) in 1.1 ml H 2 0. The rIL-2 used for die study had a specific activity of 9 x 10 6 units/mg protein. All dosages for the study were calculated as International Units (IU), using a specific activity of 15 x 10*
  • HIV-reactive antibodies if they were HIV p24 antigen negative, and if they had no concurrent opportunistic infections.
  • criteria for eligibility included an absolute CD4+ T-cell count of between 200 and 500 cell/mm 3 and being on nucleoside-analogue antiretroviral medication (e.g.
  • AZT AZT, 3TC, ddl, d4T
  • IL-2 therapy Any individual that had any chronic illness unable to be controlled by medication, pregnant or nursing, or had any history of malignancy was excluded.
  • the mean CD4+ T-cell count was 338 (range 202-495). The group was homogeneous with respect to leukocyte and differential counts, and there were no differences in these parameters among the subjects who were given different IL-2 doses. Twelve of die 14 individuals enrolled had been taking anti- retroviral nucleoside analogue medication for at least 3 months. Two subjects began anti-nucleoside medication 6 weeks prior to initiation of IL-2 therapy.
  • Example 4 Doses and Regimes for 5 Dose Groups Group 1: 125,000 rIL-2/m 2 /day
  • the administration ofthe drug was stopped in the two individuals experiencing fever, myalgia and fatigue after 4 to 5 days of inoculating 250,000 IU rIL-2/ m 2 /day, and 6 to 8 hours after each rIL-2 injection.
  • Groupe 5 500,000 IU rIL-2/mVday Four subjects received 500,000 IU (33 ⁇ g) rIL-2/m 2 /day IL-2. All the individuals treated at this dose developed systemic symptoms after 4 to 5 days of injections, and were unable to undergo further treatment at this dose level. The treatment was, thus, discontinued.
  • Toxicity was defined as two consecutive abnormal values found for any one individual.
  • Dose limiting toxicity (DLT) was defined as ⁇ WHO (World Health Organization) grade 1 toxicity, as shown in Table 2 above. If, and when, DLT occurred in any one individual, treatment was withheld from tiiat particular individual until the toxic symptoms or signs resolved. Treatment was then resumed at an IL-2 dose midway between the previous dose level and the current dose (the dose was halved). The objective was to achieve a dose tiiat was non-toxic but that would permit each subject to carry out normal daily activities.
  • Example 6 Determination of IL-2 Plasma Concentration Plasma IL-2 concentrations were determined by ELISA (Endogen, Inc. , Boston, MA), according to die manufacturer's specification. The lower limit of detection of IL-2 in plasma was 0.5 pM (7.5 pg/ml).
  • IL-2 low, but detectable levels of IL-2 were measurable in 11 of the 14 patients before treatment with IL-2 was started.
  • the mean plasma IL-2 concentration of the samples was 2.5 ⁇ 0.6 pM SEM (range 0.5 to 8.4 pM).
  • the IL-2 plasma concentration was determined at frequent intervals over die first 24 hours after each dose increment was administered. A marked increase in the detectable IL-2 plasma level was observed when following die time course at a dose of 250,000 IU/m 2 when compared with the profile obtained at a dose of 125,000 IU/m 2 . At die lower dose level, die peak IL-2 concentration attained was only about 3-4 pM. A doubling of the dose resulted in greater than 5 -fold increase in the peak IL-2 plasma concentration, to 25 pM. At these doses, concentrations of IL-2 that saturate about 50% of the high affinity receptors (about 10 pM) were detectable even 8 hours after die injection, and increased IL-2 plasma levels were still measurable 24 hours later. Thereafter, the measurement of trough IL-2 plasma levels just prior to the next dose revealed that further daily doses of IL-2 did not result in gradually increasing plasma IL-2 levels.
  • Example 8 Determination of Circulating Leukocyte Count for Individuals in First Clinical Trial
  • Example 9 Effects of IL-2 Therapy on Leukocyte Production by Individuals in First Clinical Trial
  • Linear trends in NK cell counts o ⁇ irririg during the trial period were calculated for each treatment group as a mean of the slopes obtained for each individual subject using data obtained at monthly intervals.
  • the concentration of circulating eosinophils was also observed to change.
  • An increased eosinophil count was observed after only 2 weeks of rIL-2 inoculations, and peak levels occurred at 1 month after inoculation, with a subsequent decrease toward baseline within 2 months.
  • a greater than 9-fold increase in eosinophils occurred at the latter doses after 1 month of therapy, compared with a lesser than 3-fold increase for the subjects receiving the submaximal doses.
  • CD4 + T-cells of 26 cells/mm 3 was calculated for die groups receiving the intermediate and higher doses (Groups 3 and 4).
  • Plasma HTV concentrations were determined at the beginning of the clinical trial of Examples 2 and 3 by branched chain DNA assays as described by Pachl et al. (Pachl et al. , J. Acq. Immune Defic.
  • Example 11 Effect of IL-2 Therapy on Plasma HIV Cone, in Individuals in First Clinical Trial
  • the first 6 individuals entered in the study were monitored for plasma HIV RNA levels with a branched chain DNA assay (bDNA) that had a sensitivity of 10,000 viral RNA molecules/ml. Only 2 of these individuals had detectable plasma HIV levels (14 and 20X10 3 viral RNA molecules/ml, respectively). The assays of these 2 individuals became negative after initiation of IL-2 treatment and remained so throughout die 6 months of the study.
  • bDNA branched chain DNA assay
  • the plasma HIV levels of the other 4 individuals remained negative during die 6 month study interval. Subsequent to enrollment of the first 6 individuals, a second generation bDNA assay became available that is significantly more sensitive, and has a lower limit of detection of 500 HIV RNA molecules/ml. Accordingly, the next 8 individuals enrolled were monitored using the improved bDNA assay. At the initiation of IL-2 therapy, the mean plasma HIV concentration of these 8 individuals was
  • This experiment demonstrates tiiat the present maintenance (chronic) IL-2 therapy utilized in the first trial described in Examples 2 and 3 above, may be administered to individuals for prolonged periods of time in excess of 2 years and beyond, witii extended beneficial results.
  • 10 asymptomatic HIV+ individuals are recruited to self- administer IL-2 subcutaneously, in daily doses ranging from 187,500 to 250,000 IU/m 2 (12.5-16.7 ⁇ g/m 2 ) body surface area (or 4,000 to 5,350 IU/kg body weight) for at least 12 months.
  • Subjects are eligible for study if they test positive for HIV-reactive antibodies, and if they have plasma HIV detectable by the branched-chain DNA (bDNA) assay of Pachl et al. (1995), suprs, which has a lower limit of detection of 500 RNA equivalents/ml.
  • bDNA branched-chain DNA
  • Each individual receives antiviral medication concomitantly with the IL-2 therapy consisting of zidovudine (AZT, 600 mg/d), and lamivudine (3TC, 300 mg/d) throughout the 12 month IL-2 treatment interval.
  • IL-2 therapy consisting of zidovudine (AZT, 600 mg/d), and lamivudine (3TC, 300 mg/d) throughout the 12 month IL-2 treatment interval.
  • Each individual is monitored monthly for clinical signs and symptoms of adverse effects, and for laboratory evidence of organ system dysfunction.
  • Immune system parameters studied include cutaneous tests for delayed-type- hypersensitivity (DTH) to common recall antigens, including Candida albicans, dermatophyton, trichophyton, mumps virus, and tetanus toxoid.
  • DTH delayed-type- hypersensitivity
  • WBC Circulating white blood cells
  • NK natural killer cells
  • monocytes monocytes
  • PMNs polymorphonuclear leukocytes
  • T cell subsets identified by the CD4 and CD8 surface markers are noted, along with surface markers indicative of activation, including the receptor IL-2R ⁇ chain, and IL-2R ⁇ chain, MHC Class II, CD40 Ligand (CD95), and Fas ligand.
  • Example 13 Comparison of Joint AZT IL-2 Therapy with Treatment with AZT Alone of Asymptomatic HIV+ Individuals
  • the Concorde is a double blind randomized study on the administration of AZT to asymptomatic HTV+ individuals (Th& Lancet 343: 871-881 (1994)).
  • the recruited individuals were divided into two groups, an early administration group and a second group where AZT administration was started after die individual's T-cell count fell below a certain figure or the individual developed ARC or AIDS.
  • AZT was administered at a dose of 250 mg four times per day to 877 asymptomatic HIV+ individuals and a control group of 872 individuals were given a placebo. The individuals were followed-up until death or for 3 years.
  • 418 started AZT treatment at some point in the trial, 174 after developing ARC or AIDS, and the remainder based on their low CD4 cell counts. No statistical difference was observed between the two regimes in the results observed.
  • the combination of anti-viral treatment with immunotherapy with the present low doses of IL-2 provides unexpected benefits when compared with anti-viral therapy alone. While artisans witii skill in the art have been utilizing substantially higher doses of IL-2 for therapy, the present low dose regime in combination with an anti-viral drug produces anti ⁇ viral effects with a concomitant stimulation of the immune system, in the absence of toxicity WHO Grade 1 or higher. Thus, the combined therapy precludes d e onset of immunodeficiency and the clinical diagnosis of ARC and AIDS without the toxicity observed witii other treatments.
  • CD4+ T-cells was unchanged from base line in both, the AZT-treated and in the placebo-treated, groups. On the basis of the clinical trial reported herein in Examples 2 and 3 above, the present immunotiierapy produced an increase in the concentration of circulating CD4+ T-cells of about 26 cells/mm 3 per month.
  • IU/m 2 IL-2 daily doses for self-administration when signs of a cold, fever, myalgia, cough, or rhinitis appear.
  • the individuals self-administer die IL-2 before and during the course of the infection and resolution of the symptoms of the cold or flu.
  • the individuals in Group 1 greatly benefit from the treatment in comparison with Group 2, particularly with regard to the rate of onset of symptoms, and the rate of resolution of symptoms.
  • the individuals are randomized to receive antiviral therapy vs. antiviral therapy plus IL-2 immunotherapy. 25 individuals are placed in each treatment group.
  • Antiviral therapy consists of a combination of the nucleoside analogues zidovudine (AZT,
  • lamivudine 3TC, 300 mg/day
  • protein inhibitor invirase 600 mg/day
  • Group 2 Combined Anti-viraI/IL-2 Therapy 25 individuals receive anti-viral therapy as in Group 1 plus IL-2 over the same period of time.
  • the IL-2 is self-administered daily by subcutaneous injection at 250,000 IU IL-2/m 2 (16.7 ⁇ g/m 2 ).
  • Plasma HIV levels are determined by die bDNA assay of Pachl et al. (1995), supra, and immunological tests conducted include cutaneous tests for delayed-type hypersensitivity (DTH), and white blood cell (WBC) enumeration by flow cy tome try.
  • DTH delayed-type hypersensitivity
  • WBC white blood cell
  • linear trends in cell counts and HIV plasma levels over time are calculated by computing linear regression slopes separately for each subject, and then the average slopes for each treatment group is compared using the Students' T test. Following 6 months of therapy, each treatment group is crossed-over to die alternative therapy, and the study continues for an additional 6 months.
  • Example 16 Clinical Trial with IL-2 Immunotherapy on Individuals with Congenital and Acquired Immunodeficiencies 10 individuals suffering from congenital immunodeficiencies, particularly those due to an inability to produce IL-2, and individuals suffering from acquired immunodeficiencies tiiat are not secondary to HIV infection, such as common variable immunodeficiencies, and immunodeficiencies following cytotoxic treatments for cancer (e. g., chemotherapy and radiotherapy), are recruited for IL-2 immunotherapy providing chronic immune stimulation with IL-2. These individuals self-administer IL-2 treatment protocols for 6 months to 1 year, similar to those employed for asymptomatic HIV+ individuals, shown in Example 3 above, except that they do not receive AZT.
  • cytotoxic treatments for cancer e. g., chemotherapy and radiotherapy
  • the daily dose self- injected is 250,000 IU IL-2/m 2 body surface. Noticeable improvement is noticed with respect to the initial symptomatology, including suceptibility to infections, particularly those caused by opportunistic organisms, and by a reduced requirement for other immune enhancing therapies, such as the administration of i.v. immunoglobulin.
  • Example 17 Clinical Trial with IL-2 Immunotherapy for
  • microbial infections are, for example, those due to viruses (e. g., hepatitis viruses), mycobacteria (e. g., tuberculosis, especially multi-drug-resistant M. tuberculosis), fungi (e. g., candidiasis, aspergillosis, histoplasmosis, coccidiomycosis, and nocardiosis), and parasites (e. g., toxoplasmosis, pneumocystosis), benefit from chronic immune stimulation.
  • viruses e. g., hepatitis viruses
  • mycobacteria e. g., tuberculosis, especially multi-drug-resistant M. tuberculosis
  • fungi e. g., candidiasis, aspergillosis, histoplasmosis, coccidiomycosis, and nocardiosis
  • parasites e. g.,
  • Example 19 IL-2 Therapy Administered by Other Routes This experiment has been designed to demonstrate that chronic IL-2 immunotherapy, in accordance with this invention, is beneficial without eliciting substantial toxicity WHO Grade 1 or higher, when administered by routes other than injection.
  • Example 20 Ten individuals are recruited into the study, including normal individuals, individuals afflicted with cancer, and invididuals infected with HIV and other microbial infections, and congenital and acquired immunodeficiencies. All individuals are matched with at least one corresponding individual, who self- administered IL-2 by injection under similar conditions. All individuals are instructed as described in Example 3 above to self-apply a patch or to inhale a daily dose of 250,000 IU/m 2 of IL-2 for 6 months to 1 year. All individuals in this study report on their progress, including symptoms, severity, and others, and date of occurrence, scoring on a scale of 1 to 10, from the mildest to the most severe symptoms, including fever, fatigue, myalgia, nausea, and vomiting. The results obtained are similar to those when IL-2 immunotherapy is conducted by injection.
  • Example 20 Therapy Utilizing Different Forms of the Agent
  • Ten individuals are recruited into this study, including normal individuals, individuals afflicted with cancer, HIV and odier microbial infections, and congenital and acquired immunodeficiencies.
  • a dose of derivatized IL-2 corresponding to 250,000 IU/m 2 of IL-2 is self-administered daily by subcutaneous injection by each individual over a period of 6 months to 1 year.
  • a dose of mutated IL-2 corresponding to 250,000 IU/m 2 of IL-2 is self-administered daily by each individual for a period of six months.
  • a dose of mutated X-IL-2 corresponding to 250,000 IU/m 2 of IL-2 is self administered daily by each individual for a period of six months.
  • Tests and reports are as described above. Each individual is matched witii an individual receiving recombinant IL-2 by injection as described in Example 3 above. Results which are similar to those obtained with recombinant IL-2 immunotherapy are observed.

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Abstract

Cette invention se rapporte à un procédé pour activer le système immunitaire d'un sujet, ce procédé consistant à administrer de manière chronique de faibles doses d'un agent tel que IL-2, des protéines de fusion de cet agent et des dérivés de cet agent qui sont acceptables sur le plan pharmaceutique. Cet agent est fourni sous la forme d'une composition dermique, d'un dispositif d'administration transdermique et d'un dispositif d'électro-transport, ainsi que sous la forme d'un kit en vue d'une auto-administration.
PCT/US1997/003306 1996-02-28 1997-02-26 Stimulation de reponses immunitaires au moyen de faibles doses d'interleukine-2 WO1997031622A1 (fr)

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US6955807B1 (en) 1998-05-15 2005-10-18 Bayer Pharmaceuticals Corporation IL-2 selective agonists and antagonists
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US12006354B2 (en) 2017-05-24 2024-06-11 Novartis Ag Antibody-IL2 engrafted proteins and methods of use in the treatment of cancer

Also Published As

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EP0893991A4 (fr) 2001-02-21
EP0893991A1 (fr) 1999-02-03
AU2526797A (en) 1997-09-16
US6045788A (en) 2000-04-04

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